Development of the Casparian strip is delayed by blue light in pea stems

To understand the regulatory mechanisms involved in tissue development by light, the kinetics of regulation of Casparian strip (CS) development in garden pea stems was studied. We found that short-term irradiation with white light delayed the development of the CS and used this delay to assess the quantitative effect of light on CS development. We examined the effect of the duration and fluence rates of white light treatment on CS development and observed a significant relationship between fluence and the delay in CS development indicating that the Bunsen–Roscoe law of reciprocity holds for this response. The effect of white light irradiation was not inhibited in the presence of a photosynthetic inhibitor, DCMU, or a carotenoid biosynthesis inhibitor, Norflurazon, indicating that the delay in CS development by light is a photomorphogenetic response rather than a subsidiary effect mediated by photosynthetic activity. An action spectrum for the response displayed a major peak in the blue-light region, suggesting a dominant role for blue-light receptors. A minor peak in the red-light region also suggested the possible involvement of phytochromes. Although phytochromes are known to contribute to blue-light responses, phytochrome-deficient mutants showed a normal delay of CS development in response to blue light, indicating that the response is not mediated by phytochrome and suggesting a role for one or more specific blue-light receptors.     

Rapid,Blue-Light-Induced Acidifications at the Surface of Ectocarpus and Other Marine Macroalgae

In most brown algae, photosynthesis saturated with red light can be stimulated by continuous blue light. Pulses of blue light lead to transient increases in photosynthetic rate. When a CO2-sensitive electrode was used, occasionally blue light was observed to cause an apparent increase of CO2 instead of the expected decrease. This was changed by buffering the seawater medium and, under these conditions, blue light caused stimulation of CO2 consumption. These results led to investigations of blue-light-dependent pH changes at the outer surface of the plants. Shifts of the pH were recorded in the presence of the photosynthetic inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea. In all brown algae tested and in the green algae Ulva and Enteromorpha, blue-light pulses caused transient acidification of 0.03 to 0.18 pH units, depending on the species. The kinetics showed lag phases of a few seconds and the minimum was reached after 5 to 9 min. Fluence response relationships indicated that the sensitivity (threshold) to blue light was very similar in all species. The responses in Ectocarpus changed with time, and about 5 h after the beginning of red light or darkness, a second component became evident, which peaked 20 min after the blue-light pulse. The refractory period of the whole system was about 3 h in Ectocarpus. The blue-light-dependent pH changes show striking similarities to those of higher plant guard cells, and it is possible that similar responses may occur in other tissues of higher plants. In red algae, however, no blue-light-dependent acidifications could be detected. The possible role of the observed pH shifts in a mechanism of CO2 acquisition is discussed.     

KINETICS OF BLUE-LIGHT STIMULATION AND CIRCADIAN RHYTHMICITY OF LIGHT-SATURATED PHOTOSYNTHESIS IN BROWN ALGAE: A SPECIES COMPARISON1

The time courses of photosynthetic rates in red light, with and without additional blue light, were investigated and compared in 20 species of brown algae. Species could be separated into two groups on the basis of the rhythmicity of their photosynthesis in red light and the kinetics of their responses to blue-light pulses. One group, which consisted of members of the Ectocarpales, Chordariales, and Dictyosiphonales, was characterized by strong and persistent circadian rhythmicity in red light. The photosynthetic responses of these species to blue-light pulses started within 10–30 s of the beginning of blue-light treatment and mostly contained at least two distinct kinetic components. An early component, which reached a maximum about 5–10 min after the blue-light pulse, was always detectable. Later components were seen as separate peaks or shoulders after an additional 10–20 min. The decay of the response in this group of species was mostly slow, with half-lives of between 0.5 and 1.5 h. In the second group of species, consisting of members of the Dictyotales, Laminariales, and Fucales, photosynthesis in red light was usually non-rhythmic, although circadian rhythms with a weak amplitude or of transient occurrence were observed in some plants of some species. The increase in photosynthesis in response to a blue-light pulse was not detectable until 70–330 s after the start of blue-light treatment, and the response itself had only a single component, with a maximum after about 10 min and half-life of 10–20 min. The lengths of the lag-phases were positively correlated with the times taken to reach the peak in this group, although the lag-phases and the half lives sometimes varied with time in individual plants. Two members of the Sphacelariales (Sphacelaria, Cladostephus) did not fit into either of the two groups because their photosynthesis was rhythmic, but their responses had long lag-phases, a single component, and moderately long half-lives. The differences in the kinetics of the photosynthetic response to blue-light pulses, which have been described for the two main groups of species, are thought to indicate that there are two distinct mechanisms by which light-saturated photosynthesis responds to blue light in brown algae. Since in some species the maximal photosynthesis after a blue-light pulse and the rate of photosynthesis in continuous blue light also varied in a circadian pattern, the response to blue light itself may be under circadian control.     

Genome-wide analysis of light sensing in Prochlorococcus

Prochlorococcus MED4 has, with a total of only 1,716 annotated protein-coding genes, the most compact genome of a free-living photoautotroph. Although light quality and quantity play an important role in regulating the growth rate of this organism in its natural habitat, the majority of known light-sensing proteins are absent from its genome. To explore the potential for light sensing in this phototroph, we measured its global gene expression pattern in response to different light qualities and quantities by using high-density Affymetrix microarrays. Though seven different conditions were tested, only blue light elicited a strong response. In addition, hierarchical clustering revealed that the responses to high white light and blue light were very similar and different from that of the lower-intensity white light, suggesting that the actual sensing of high light is mediated via a blue-light receptor. Bacterial cryptochromes seem to be good candidates for the blue-light sensors. The existence of a signaling pathway for the redox state of the photosynthetic electron transport chain was suggested by the presence of genes that responded similarly to red and blue light as well as genes that responded to the addition of DCMU [3-(3,4-dichlorophenyl)-1,1-N-N'-dimethylurea], a specific inhibitor of photosystem II-mediated electron transport.     

Photoresponses in Rhodobacter sphaeroides: role of photosynthetic electron transport.

Rhodobacter sphaeroides responds to a decrease in light intensity by a transient stop followed by adaptation. There is no measurable response to increases in light intensity. We confirmed that photosynthetic electron transport is essential for a photoresponse, as (i) inhibitors of photosynthetic electron transport inhibit photoresponses, (ii) electron transport to oxidases in the presence of oxygen reduces the photoresponse, and (iii) the magnitude of the response is dependent on the photopigment content of the cells. The photoresponses of cells grown in high light, which have lower concentrations of light-harvesting photopigment and reaction centers, saturated at much higher light intensities than the photoresponses of cells grown in low light, which have high concentrations of light-harvesting pigments and reaction centers. We examined whether the primary sensory signal from the photosynthetic electron transport chain was a change in the electrochemical proton gradient or a change in the rate of electron transport itself (probably reflecting redox sensing). R. sphaeroides showed no response to the addition of the proton ionophore carbonyl cyanide 4-trifluoromethoxyphenylhydrazone, which decreased the electrochemical proton gradient, although a behavioral response was seen to a reduction in light intensity that caused an equivalent reduction in proton gradient. These results strongly suggest that (i) the photosynthetic apparatus is the primary photoreceptor, (ii) the primary signal is generated by a change in the rate of electron transport, (iii) the change in the electrochemical proton gradient is not the primary photosensory signal, and (iv) stimuli affecting electron transport rates integrate via the electron transport chain.     

A single flavoprotein,AppA, integrates both redox and light signals in Rhodobacter sphaeroides

Anoxygenic photosynthetic proteobacteria exhibit various light responses, including changing levels of expression of photosynthesis genes. However, the underlying mechanisms are largely unknown. We show that expression of the puf and puc operons encoding structural proteins of the photosynthetic complexes is strongly repressed by blue light under semi-aerobic growth in Rhodobacter sphaeroides but not in the related species Rhodobacter capsulatus. At very low oxygen tension, puf and puc expression is independent of blue light in both species. Photosynthetic electron transport does not mediate the blue light repression, implying the existence of specific photoreceptors. Here, we show that the flavoprotein AppA is likely to act as the photoreceptor for blue light-dependent repression during continuous illumination. The FAD cofactor of AppA is essential for the blue light-dependent sensory transduction of this response. AppA, which is present in R. sphaeroides but not in R. capsulatus, is known to participate in the redox-dependent control of photosynthesis gene expression. Thus, AppA is the first example of a protein with dual sensing capabilities that integrates both redox and light signals.     

Circadian Rhythms in Stomatal Responsiveness to Red and Blue Light

Stomata of many plants have circadian rhythms in responsiveness to environmental cues as well as circadian rhythms in aperture. Stomatal responses to red light and blue light are mediated by photosynthetic photoreceptors; responses to blue light are additionally controlled by a specific blue-light photoreceptor. This paper describes circadian rhythmic aspects of stomatal responsiveness to red and blue light in Vicia faba. Plants were exposed to a repeated light:dark regime of 1.5:2.5 h for a total of 48 h, and because the plants could not entrain to this short light:dark cycle, circadian rhythms were able to "free run" as if in continuous light. The rhythm in the stomatal conductance established during the 1.5-h light periods was caused both by a rhythm in sensitivity to light and by a rhythm in the stomatal conductance established during the preceding 2.5-h dark periods. Both rhythms peaked during the middle of the subjective day. Although the stomatal response to blue light is greater than the response to red light at all times of day, there was no discernible difference in period, phase, or amplitude of the rhythm in sensitivity to the two light qualities. We observed no circadian rhythmicity in net carbon assimilation with the 1.5:2.5 h light regime for either red or blue light. In continuous white light, small rhythmic changes in photosynthetic assimilation were observed, but at relatively high light levels, and these appeared to be attributable largely to changes in internal CO2 availability governed by stomatal conductance.     

Acclimation of Arabidopsis thaliana to the light environment: regulation of chloroplast composition

The regulation by light of the composition of the photosynthetic apparatus was investigated in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta. When grown in high- and low-irradiance white light, wild-type plants and photomorphogenic mutants showed large differences in their maximum photosynthetic rate and chlorophyll a/b ratios; such changes were abolished by growth in red light. Photosystem I (PSI) and PSII levels were measured in wild-type plants grown under a range of light environments; the results indicate that regulation of photosystem stoichiometry involves the specific detection of blue light. Supplementing red growth lights with low levels of blue light led to large increases in PSII content, while further increases in blue irradiance had the opposite effect; this latter response was abolished by the hy4 mutation, which affects certain events controlled by a blue-light receptor. Mutants defective in the phytochrome photoreceptors retained regulation of photosystem stoichiometry. We discuss the results in terms of two separate responses controlled by blue-light receptors: a blue-high-fluence response which controls photosystem stoichiometry; and a blue-low-fluence response necessary for activation of such control. Variation in the irradiance of the red growth light revealed that the blue-high-fluence response is attenuated by red light; this may be evidence that photosystem stoichiometry is controlled not only by photoreceptors, but also by photosynthetic metabolism.Abbreviations BHF blue-high-fluence - BLF blue-low-fluence - Chl chlorophyll - FR far-red light - LHCII light-harvesting complex of PSII - P_max maximum photosynthetic rate - R red light - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase This work was supported by Natural Environment Research Council Grant No. GR3/7571A. We would like to thank H. Smith (Botany Department, University of Leicester) and E. Murchie (INRA, Versailles) for helpful discussions.     

Evidence that the long-lifetime photointermediate of s-rhodopsin is a receptor for negative phototaxis in Halobacterium halobium

The effect of blue background light on behavioral response of Halobacterium halobium to step-like stimulation with green-orange attractant light was examined. The results strongly support the previously proposed hypothesis that a long-lifetime photointermediate of s-rhodopsin is the photoreceptor for repellent light: the step-like increase in green-orange light was convertible from attractant stimulus to repellent one, when the cells were constantly illuminated with blue light. No difference of the threshold intensity of the blue background light was observed between the mutant strain that lacks both bacteriorhodopsin and halorhodopsin and the wild type strain, suggesting that the two light-driven ion pumps are not participant in sensing attractant light.     

Reorientation of microtubules at the outer epidermal wall of maize coleoptiles by phytochrome,blue-light photoreceptor,and auxin

Summary The effects of red and blue light on the orientation of cortical microtubules (MTs) underneath the outer epidermal wall of maize (Zea mays L.) coleoptiles were investigated with immunofluorescent techniques. The epidermal cells of dark-grown coleoptiles demonstrated an irregular pattern of regions of parallel MTs with a random distribution of orientations. This pattern could be changed into a uniformly transverse MT alignment with respect to the long cell axis by 1 h of irradiation with red light. This response was transient as the MTs spontaneously shifted into a longitudinal orientation after 1–2 h of continued irradiation. Induction/reversion experiments with short red and far-red light pulses demonstrated the involvement of phytochrome in this response. In contrast to red light, irradiation with blue light induced a stable longitudinal MT alignment which was established within 10 min. The blue-light response could not be affected by subsequent irradiations with red or far-red light indicating the involvement of a separate blue-light photoreceptor which antagonizes the effect of phytochrome. In mixed light treatments with red and blue light, the blue-light photoreceptor always dominated over phytochrome which exhibited an apparently less stable influence on MT orientation. Long-term irradiations with red or blue light up to 6 h did not reveal any rhythmic changes of MT orientation that could be related to the rhythmicity of helicoidal cell-wall structure. Subapical segments isolated from dark-grown coleoptiles maintained a longitudinal MT arrangement even in red light indicating that the responsiveness to phytochrome was lost upon isolation. Conversely auxin induced a transverse MT arrangement in isolated segments even in blue light, indicating that the responsiveness to blue-light photoreceptor was eliminated by the hormone. These complex interactions are discussed in the context of current hypotheses on the functional significance of MT reorientations for cell development.Abbreviations MT cortical microtubule - P_r, P_fr red and far-red absorbing form of phytochrome     

Electron transport-dependent taxis in Rhodobacter sphaeroides.

Rhodobacter sphaeroides showed chemotaxis to the terminal electron acceptors oxygen and dimethyl sulfoxide, and the responses to these effectors were shown to be influenced by the relative activities of the different electron transport pathways. R. sphaeroides cells tethered by their flagella showed a step-down response to a decrease in the oxygen or dimethyl sulfoxide concentration when using them as terminal acceptors. Bacteria using photosynthetic electron transport, however, showed a step-down response to oxygen addition. Addition of the proton ionophore carbonyl cyanide 4-trifluoromethoxyphenylhydrazone did not cause a transient behavioral response, although it decreased the electrochemical proton gradient (delta p) and increased the rate of electron transport. However, removal of the ionophore, which caused an increase in delta p and a decrease in the electron transport rate, resulted in a step-down response. Together, these data suggest that behavioral responses of R. sphaeroides to electron transport effectors are caused by changes in the rate of electron transport rather than changes in delta p.     

不产氧光合细菌光合色素的光氧调控机制

[目的]为不产氧光合细菌光合色素研究提供可行的较系统规范的研究方法和数据,揭示固氮红细菌(Rhodobacter azotoformans 134K20)光合色素光氧适应性机制.[方法]采用光谱法和色谱法对光和氧调控下的类胡萝卜素和细菌叶绿素合成代谢进行了研究.[结果]134K20菌株光照好氧时细胞得率最高.光照厌氧时主要合成3黄、1红、1紫、2绿、2蓝9种色素,黄色素大量表达.有氧时红色素大量表达,且启动2种新的红色素和1种新的紫色素表达,而黄色和蓝绿色素则受氧抑制.黑暗好氧主要合成2黄、3红、2紫、1绿、1蓝9种色素,但不同于光照厌氧.光照好氧时黄色素减少到1种,紫色素含量增加,其余同黑暗好氧.[结论]固氮红细菌(Rhodobacter azotoformans 134K20)是通过PpsR调节途径来调节光合基因表达的.黄色和红色素属于类胡萝卜素.黄色素1属于球形烯系列,其余两种黄色素是新的类胡萝卜素组分.红色素为新的球形烯酮组分,3种红色素极性、峰形和峰位差别显著,正己烷能显示其精细结构.紫色为极性较大的细菌脱镁叶绿素,绿色和蓝色为4种极性不同的细菌叶绿素a中间产物.乙醚甲醇法适合类胡萝卜素的提取,丙酮甲醇冰冻研磨法能快速有效完全提取光合色素.溶剂效应可有效鉴别细菌叶绿素a中间产物.     

Redox and light regulation of gene expression in photosynthetic prokaryotes

All photosynthetic organisms control expression of photosynthesis genes in response to alterations in light intensity as well as to changes in cellular redox potential. Light regulation in plants involves a well-defined set of red- and blue-light absorbing photoreceptors called phytochrome and cryptochrome. Less understood are the factors that control synthesis of the plant photosystem in response to changes in cellular redox. Among a diverse set of photosynthetic bacteria the best understood regulatory systems are those synthesized by the photosynthetic bacterium Rhodobacter capsulatus. This species uses the global two-component signal transduction cascade, RegB and RegA, to anaerobically de-repress anaerobic gene expression. Under reducing conditions, the phosphate on RegB is transferred to RegA, which then activates genes involved in photosynthesis, nitrogen fixation, carbon fixation, respiration and electron transport. In the presence of oxygen, there is a second regulator known as CrtJ, which is responsible for repressing photosynthesis gene expression. CrtJ responds to redox by forming an intramolecular disulphide bond under oxidizing, but not reducing, growth conditions. The presence of the disulphide bond stimulates DNA binding activity of the repressor. There is also a flavoprotein that functions as a blue-light absorbing anti-repressor of CrtJ in the related bacterial species Rhodobacter sphaeroides called AppA. AppA exhibits a novel long-lived photocycle that is initiated by blue-light absorption by the flavin. Once excited, AppA binds to CrtJ thereby inhibiting the repressor activity of CrtJ. Various mechanistic aspects of this photocycle will be discussed.     

Transient reduction of responsiveness of blue-light-mediated hair-whorl morphogenesis inAcetabularia mediterranea induced by blue light

After a prolonged period of red light the formation of a new whorl of lateral hairs can be induced inAcetabularia mediterranea Lamouroux (=A. acetabulum (L.) Silva) by a pulse of blue light. It has previously been shown that the response to blue light obeys the law of reciprocity. In this paper we demonstrate that the responses to blue light are additive only within 10 min after the onset of blue-light treatment, since the responsiveness of the cells is also affected by blue light. One hour after a short blue-light pulse the response to a second blue-light pulse has come to a minimum. After that, the responsiveness is restored in a refractory period of several hours. The fluenceresponse curves for hair-whorl formation at the time of minimum responsiveness are shifted parallel to the original fluence-response curves without preirradiation. Again, the law of reciprocity applies. This indicates an increased light requirement only for the same degree of hair-formation response. The sensitivity to blue light of the reduction of responsiveness response is higher by a factor of about 50 than the induction of hairformation response.     

Blue-Light-Induced Shrinking of Protoplasts from Maize Coleoptiles and Its Relationship to Coleoptile Growth

Protoplasts isolated from red-light-grown maize (Zea mays L.) coleoptiles shrank transiently upon brief exposure (e.g. 30 s) to blue light under background irradiation with red light. The maximal volume reduction (about 4% at a saturating fluence) occurred about 5 min after blue-light stimulation. The response was prevented by the anion-channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid. Red light and far-red light did not induce any comparable response. Protoplasts of different sizes and those isolated from different coleoptile positions showed similar responses. After treatment with a saturating blue-light pulse, the protoplasts became responsive to a second pulse and gained full responsiveness within 5 min, suggesting that the photoreceptor system involves a dark-reversible component. The response to continuous blue light was also found to be transient. The protoplast volume was reduced during about 6 to 9 min of irradiation and returned within the next 30 min to the control level. The response to continuous blue light was saturated at 30 [mu]mol m-2 s-1. However, when the fluence rate was enhanced 10-fold after a period of irradiation at 30 [mu]mol m-2 s-1, the protoplasts showed another shrinking response. These and other kinetic results indicate that the photoreceptor system undergoes a photosensory adaptation. Growth in different zones of the coleoptile was inhibited by blue light transiently after pulse stimulation, as well as during continuous stimulation. It was concluded that the observed protoplast shrinking is related to the blue-light-induced inhibition of coleoptile growth.     

Evidence for two different blue-light-receptor systems for the fast responses of stimulation of photosynthetic capacity and acidification of the plant surface in brown algae

Two blue-light responses of Phaeophyta that are expressed within a few seconds of a blue-light stimulus were characterized with respect to their photoreception properties. The first response is the activation of red-light-saturated photosynthesis which can be stimulated to values up to 5 times the rates in red light, depending on the species. The second response is a blue-light-induced acidification measurable at the plant surface. Both responses have similar kinetic characteristics and thus led us initially to hypothesise that they were causally connected in the same transduction mechanism. The two responses have action spectra [measured for Ectocarpus siliculosus (Dillwyn) Lyngb. and Laminaria saccharina (L.) Lamouroux] that are indistinguishable within the relatively large limits of error. However, in all species tested, the threshold sensitivity for blue light of the photosynthetic response is lower than that of the pH-shift by a factor of 2 to 150. Furthermore, stimulation of photosynthesis is sensitive to the flavin inhibitors, KI and phenylacetic acid, but the pH response is not affected by these inhibitors. Thus, the blue-light-induced pH-shift does not cause the stimulation of photosynthesis. In contrast, the different fluence-response relationships of the two responses and particularly the differential effect of the inhibitors are clear evidence for the action of two independent transduction pathways and photoreceptor systems for blue light. At least photoreception for stimulation of photosynthesis involves a flavin-or and a pterin.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - PAA phenylacetic acid We thank Dr. C. A. Maggs for collecting P. pavonica. This research was supported by National Environment Research Council grant No. GR3/8102.     

The eubacterium Ectothiorhodospira halophila is negatively phototactic, with a wavelength dependence that fits the absorption spectrum of the photoactive yellow protein.

The motile, alkalophilic, and extremely halophilic purple sulfur bacterium Ectothiorhodospira halophila is positively photophobotactic. This response results in the accumulation of bacteria in light spots (E. Hustede, M. Liebergesell, and H. G. Schlegel, Photochem. Photobiol. 50:809-815, 1989; D. E. McRee, J. A. Tainer, T. E. Meyer, J. Van Beeumen, M. A. Cusanovich, and E. D. Getzoff, Proc. Natl. Acad. Sci. USA 86:6533-6537, 1989; also, this work). In this study, we demonstrated that E. halophila is also negatively phototactic. Video analysis of free-swimming bacteria and the formation of cell distribution patterns as a result of light-color boundaries in an anaerobic suspension of cells revealed the existence of a repellent response toward intense (but nondamaging) blue light. In the presence of saturating background photosynthetic light, an increase in the intensity of blue light induced directional switches, whereas a decrease in intense blue light gave rise to suppression of these reversals. To our knowledge, this is the first report of a true repellent response to light in a free-swimming eubacterium, since the blue light response in Escherichia coli and Salmonella typhimurium (B. L. Taylor and D. E. Koshland, Jr., J. Bacteriol. 123:557-569, 1975), which requires an extremely high light intensity, is unlikely to be a sensory process. The wavelength dependence of this negative photoresponse was determined with narrow band pass interference filters. It showed similarity to the absorption spectrum of the photoactive yellow protein from E. halophila.